Effect of Direct-Fed Microbials and Enzyme Supplementation in Prepartum Holstein Cows on Colostrum and Calf Immunity

In cows, colostrum is composed of several antibodies and nutrients to provide immunity and energy to the calf. Feeding calves high quality colostrum has been shown to improve calf health, leading to reduced mortality in calves and greater milk production in cows. The addition of direct-fed microbials (DFM) to cow diets has been theorized to improve feed efficiency and milk production, with studies showing mixed results. However, few experiments have studied the effect of feeding DFM on colostrum quality. In this experiment two treatments were given, 1) DFM and 2) DFM and enzymes (DFME). Colostrum was analyzed to determine if yield, composition, and immunoglobulin (IgG and IgA) concentration were affected. Calf serum IgG and IgA concentrations were analyzed to determine if 24 h concentrations and apparent efficiency of absorption (AEA) were affected. There were no differences with regard to yield or IgA concentration. The percent of ash showed a positive trend, indicating a higher percentage with the treatments (P = 0.067). The treatments had no effect on the additional components analyzed. The results for the IgG concentration were not significant although an increase was observed from 79.1 mg/mL in the control to 91.1 mg/mL in the DFME treatment. Neither treatment had an effect on calf immunoglobulin concentration or AEA. Based on the results, feeding DFM or DFME improves percent ash and might increase IgG concentration, but further research is necessary

Local Democracy and Education Policy in Newly Federal Nepal

In 2017, Nepal held its first local elections in twenty years. These were the first elections held under Nepal’s new constitution, ratified in 2015, which transitioned the country from a unitary state to a Federal Democratic Republic. This case study analyzes the effect of the transition to federalism on decision-making and community representation in local governance in Annapurna Rural Municipality in West-Central Nepal. This study focuses specifically on education policy, one of the more public and contentious policy responsibilities devolved from the federal level to the local units under the new constitution. This research is based on interviews with government officials, teachers, and other stakeholders in governance and education policy. It finds that although there is general support and optimism for the federal transition, there remain many challenges to implementation, especially in terms of lack of capacity and the persistence of old institutions. These challenges limit the extent to which the constitution’s goals of local democracy and inclusive government can be realized, especially in rural and mountainous areas of Nepal

The effect of Cisplatin on Human Diploid Fibroblast Cells expressing SV40 T-Antigen

Cancer is most commonly thought of as the result of accumulation of cellular DNA mutations that inhibit tumor suppressor proteins or activate oncogenes. However a large body of evidence also shows that the wild type gene transcripts are alternatively spliced to create oncogenic protein isoforms or induce nonsense mediated decay of the RNA. To combat the cancerous cell growth, chemotherapy targets cellular activities such as DNA replication, spindle formation, receptor signalling, and metabolism. Knowing the protein profile of the tumor is relevant to finding an effective chemotherapeutic. For instance, cisplatin is used to treat a variety of cancers but has been shown to have reduced efficacy if the cell lacks the protein BIN1 or if it is alternatively spliced to the BIN1+12A isoform. Cisplatin forms platinum-DNA adducts that create single stranded breaks, and if not repaired can later become double stranded breaks following DNA replication. Unlike other tumor suppressors, BIN1 appears to induce cell death by increasing genomic instability. BIN1 is proposed to function by binding to the transcription factor E2F1, and preventing the activation of the DNA repair mechanism by ATM. This allows the breaks produced by cisplatin to disrupt cellular processes, such as DNA replication and transcription, and ultimately results in cell death (Folk 2019). While studying the mechanism of cellular transformation by the simian virus 40 (SV40) T-antigen oncoprotein, our lab has discovered a novel RNA isoform of BIN1. This transcript retains exon 11 yet deletes exons 12-18. Concomitant with characterizing the protein structure, this study is investigating the functional characteristic of this BIN1 isoform as it relates to cisplatin resistance. Human diploid fibroblasts immortalized with telomerase HDF (HDF(tert)) and HDF(tert) cells expressing the viral oncoprotein SV40 T antigen (HDF(tert)+T) will be used as the experimental model. A dose curve of cisplatin from 0-2ug/ml will be used to assess cellular sensitivity. Determining the function of this novel BIN1 isoform is essential to the chemotherapeutic success in cancer treatment

Systemic effects of mastitis in the dairy cow : identifying mechanisms underlying impaired lactation in non-inflamed glands

Efficient milk production is temporarily compromised during the inflammatory response to lipopolysaccharide (LPS). Generally, the most severe losses are localized to individual mammary glands with overt cases of mastitis. However, systemic inflammation arising from an inflamed gland can negatively impact lactation in healthy neighboring glands. This interdependence between glands highlights the importance of systemic inflammation in regulating lactation, yet the underlying mechanisms and mediators remain poorly understood. To determine these systemic mechanisms, paired cows were randomly assigned to LPS treatment (T) or saline control (C). Within each treatment, two ipsilateral quarters received one of 2 intramammary sub-treatments: in T cows, infusion of 50 [micrograms] LPS (T-L) or saline (T-S); in C cows, infusion of saline (C-S) or no infusion (C-N). Front quarters were sampled for milk production and composition while rear quarters were biopsied at 0, 3, and 12 h, relative to infusions. Mammary tissue from 3 and 12 h biopsies was then subjected to RNA sequencing. Results from milk confirmed that LPS induced a characteristic immune response, with a local increase in milk somatic cells and total protein concentrations only in T-L quarters. In comparison, components associated with lactation were affected in both T-L and T-S quarters, with concentrations of milk fat and lactose being lower by 3 and 12 h post-infusion, respectively. Further, milk yields declined steadily over 24 h with similar losses in both T-L and T-S quarters. Induction of transient fever at 3 h, followed by changes in plasma antioxidant capacity and glucose concentrations, provided additional evidence of systemic responses to inflammation. Collectively, these results confirmed that localized mastitis affected neighboring glands and demonstrated that each milk component was affected by unique time- and treatment- dependent effects. Analyzing changes in the mammary transcriptome provided further insight into the regulation of the immune response and lactation during mastitis. Most genes were differentially expressed (DE) only in T-L glands and were associated with pro-inflammatory, cell signaling, and metabolic pathways. There was no direct link between expression of lactation-specific genes and hypogalactia or altered milk composition; however, genes unrelated to lactation were upregulated in T-S glands, suggesting that differential expression of other genes may have impacted lactation. At 3 h, the upregulation of negative feedback inhibitors of pro-inflammatory pathways in T-S tissue indicated unidirectional signaling from T-L to T-S glands, likely mediated by cytokines. Later, at 12 h, increased expression in T-S quarters of genes linked to one carbon metabolism and glucocorticoid stimulation indicated that other mediators may also impact milk production. From 3 to 12 h, expression of immune genes diminished in both glands despite increasing somatic cells in T-L quarters. Given the similar decline in quarter milk yields through 24 h, results suggested that an acute, transient inflammatory signal was sufficient to induce delayed effects on lactation. In summary, localized mastitis impaired lactation in non-inflamed, neighboring quarters through both direct and indirect actions of pro-inflammatory mediators.Includes bibliographical references (pages 88-102

Identifying instructional practices employed by Massachusetts Special Olympics Hall of Fame coaches

Thesis (Ed.D.)--Boston UniversitySpecial Olympics athletes may experience unique learning needs not seen among many mainstream athletes. Consequently, research-based coaching practices recommended for coaches of mainstream teams may not be appropriate for Special Olympics coaches. With this in mind, the importance of establishing a research-based knowledge bank of coaching practices to use with athletes who have intellectual disabilities comes to light. Identified coaching practices can benefit coaches of Special Olympics teams, as well as coaches of mainstream teams who may work with athletes who have intellectual disabilities. This first-of-its-kind empirical research study sought to identify instructional practices demonstrated by Massachusetts Special Olympics Hall of Fame coaches (n = 8). Specifically, this study aimed to discover what instructional practices were most commonly used among the participant group; what these instructional practices looked like; and what instructional practices may have contributed to supporting a positive learning climate. A framework for instructional practices set by the National Council for Accreditation of Coach Education (NCACE) was used to assist with the identification of instructional behaviors employed by study participants. A second purpose of this study was to use the NCACE (2006) Domain Five Observation Instrument (DFOI), a competency-based observation tool developed for this study, to collect data on study participants' instructional practices. Each study participant was observed coaching a practice three times during their sport season. Using the DFOI, observed instructional behaviors were quantified via event recording procedures and described by recording descriptive field notes. To triangulate data collected on the DFOI, coaches were audio-recorded with a wireless lapel microphone during practice observations. Following each coach's third practice observation, audio-recorded interviews were conducted to gain further insight into demonstrated instructional practices. Participants were observed performing 2,157 instructional behaviors during 1,080 minutes of observation, averaging 1.99 behaviors per minute and 89.87 behaviors per practice. Based on data analysis, encouragement, positive correction, and tactical and technical cues were the three instructional practices most commonly employed by study participants, accounting for 68.33% of all recorded behaviors. Data also show that participants engaged athletes in sport-specific activities during 63.15% of the total observation time, and spent only 5.95% of the total observation time delivering instruction to their teams. While speculative, data suggest that participants' emphasis of initiative, choice, excellence, and interpersonal relationships may have contributed to the support of positive learning environments. Additionally, the DFOI proved to be a useful observation tool for collecting competency-based quantitative and qualitative data, and is recommended for use during future replications of the current study, as well as future assessments of sport coaches' instructional practices. Based on data collected in this study, 14 recommendations are presented for Special Olympics, community-based, and school-based coach education programs. These recommendations include: promoting the importance of weaving encouragement into coaching practices, using a feedback model to facilitate delivery of positive corrections, and prioritizing use of instructional tools using a "tools in hand, tools in the tool belt, and tools in the toolbox" model

Brief for the Weymouth School Committee (Weymouth, Massachusetts), as Amici Curiae supporting respondents, Algonquin Gas Transmission, LLC and Maritimes & Northeast Pipeline, LLC v. Fore River residents against the compressor station, et al.

Published versio

Determining Differential Expression Patterns of MicroRNAs in Mammalian Skin by In Situ Hybridization

MicroRNAs are small non-coding RNAs that regulate gene expression posttranscriptionally (Bartel, 2009). Several different microRNAs have been implicated in regulatory pathways of skin morphogenesis, stem cells, skin cancer, and other skin diseases. A subset of microRNAs have been found to be differentially expressed in distinct skin lineages, namely epidermis and hair follicle (Yi et al., 2006). Characterization of the spatial and temporal expression patterns of these microRNAs can provide a basis to investigate their functions during development as well as in skin diseases including skin cancer. During my thesis study, I employed in situ hybridization to characterize the differential expression of four discrete microRNAs: miR-203, miR-143, miR-125b, and miR-205 in mammalian skin. My results demonstrated that miR-203 is highly expressed in differentiating skin lineages, miR-143 is enriched in the transient amplifying cells in hair follicle matrix, miR-125b and miR-205 are enriched in skin stem cells. I also cloned three regions upstream of miR-205 and analyzed their promoter activity in cultured keratinocytes. Together, my study shows that miRNAs are differentially expressed in distinct skin lineages and provides a basis to analyze their functions in the future

Energy Discovery-Innovation Institutes: A Step Toward America's Energy Sustainability

Part of the "Blueprint for American Prosperity"http://deepblue.lib.umich.edu/bitstream/2027.42/88629/1/2009_Brookings_Energy_Report.pd

Formulation and Evaluation of Cephalexin Extended Release Matrix Tablets Using 32 Factorial Design

The aim of the present investigation was to prepare extended release film coated matrix tablets of cephalexin using binary mixture of two grades of hydrophilic polymer, hydroxypropyl methyl cellulose (HPMC), by direct compression method. Results of the preliminary trials indicated that the polymers used have significant release retarding effect on the formulation. To study the effect of concentration of polymers on drug release from matrix tablets, 32 full factorial design was applied. The concentration of HPMC K15M and HPMC 15cps were used as independent variables, while percentage drug release was selected as dependent variable. The dissolution data were fitted into zero-order, first-order, Higuchi and Korsemeyer–Peppas models to identify the pharmacokinetics and mechanism of drug release. Comparative study of dissolution profile of final batch F3 with market preparation (Sporidex AF 375) was done by similarity factor (f2) determination and it was concluded that final formulation F3 (10% HPMC K15M, 17.5% HPMC 15cps) shows good similarity with the market product. The results of the accelerated stability study of final formulation F3 for 1 month revealed that storage conditions were not found to have made any significant changes in final formulation F3. The release of cephalexin was prolonged for 6 h by using polymer combinations of HPMC and a twice daily matrix tablet was formulated